Selection of Conditions for Cellulase and Xylanase Extraction from Switchgrass Colonized by Acidothermus cellulolyticus

• Published in: Applied biochemistry and biotechnology 2011-07, Vol.164 (6), p.793-803
• Main Authors: Rezaei, Farzaneh, Joh, Lawrence D., Kashima, Hiroyuki, Reddy, Amitha P., VanderGheynst, Jean S.
• Format: Article
• Language: English
• Subjects: Acidothermus cellulolyticus; Actinomycetales; Actinomycetales - chemistry; Actinomycetales - enzymology; Actinomycetales - metabolism; Bacteria; Bacterial Proteins; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Biochemistry; Biological and medical sciences; Biotechnology; buffers; Cellulase; Cellulase - isolation & purification; Cellulase - metabolism; Chemical Fractionation; Chemical Fractionation - methods; Chemistry; Chemistry and Materials Science; endo-1,4-beta-glucanase; Enzymes; enzymology; ethylene glycol; Fermentation; Fundamental and applied biological sciences. Psychology; isolation & purification; metabolism; methods; Methods. Procedures. Technologies; Microbial engineering. Fermentation and microbial culture technology; microbiology; Panicum; Panicum - chemistry; Panicum - microbiology; Panicum virgatum; sodium acetate; Sodium chloride; solid state fermentation; Xylan Endo-1,3-beta-Xylosidase; Xylan Endo-1,3-beta-Xylosidase - isolation & purification; Xylan Endo-1,3-beta-Xylosidase - metabolism; xylanases
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-011-9174-6

Solid-state fermentation has been widely used for enzyme production. However, secreted enzymes often bind to the solid substrate preventing their detection and recovery. A series of screening studies was performed to examine the role of extraction buffer composition including NaCl, ethylene glycol, sodium acetate buffer, and Tween 80, on xylanase and cellulase recovery from switchgrass. Our results indicated that the selection of an extraction buffer is highly dependent on the nature and source of the enzyme being extracted. While a buffer containing 50 mM sodium acetate at pH 5 was found to have a positive effect on the recovery of commercial fungal-derived cellulase and xylanase amended to switchgrass, the same buffer had a significant negative effect on enzyme extraction from solid fermentation samples colonized by the bacterium Acidothermus cellulolyticus . Xylanase activity was more affected by components in the extraction buffers compared to cellulase. This study demonstrated that extraction followed by diafiltration is important for assessing enzyme recovery from solid fermentation samples. Reduction in activity due to compounds present in the switchgrass extracts is reversible when the compounds are removed via diafiltration.